title: Ex vivo intracoronary gene transfer of adeno-associated virus 2 leads to superior transduction over serotypes 8 and 9 in rat heart transplants.
abstract: Heart transplant gene therapy requires vectors with long-lasting gene expression, high cardiotropism, and minimal pathological effects. Here, we examined transduction properties of ex vivo intracoronary delivery of adeno-associated virus (AAV) serotype 2, 8, and 9 in rat syngenic and allogenic heart transplants. Adult Dark Agouti (DA) rat hearts were intracoronarily perfused ex vivo with AAV2, AAV8, or AAV9 encoding firefly luciferase and transplanted heterotopically into the abdomen of syngenic DA or allogenic Wistar-Furth (WF) recipients. Serial in vivo bioluminescent imaging of syngraft and allograft recipients was performed for 6 months and 4 weeks, respectively. Grafts were removed for PCR-, RT-PCR, and luminometer analysis. In vivo bioluminescent imaging of recipients showed that AAV9 induced a prominent and stable luciferase activity in the abdomen, when compared with AAV2 and AAV8. However, ex vivo analyses revealed that intracoronary perfusion with AAV2 resulted in the highest heart transplant transduction levels in syngrafts and allografts. Ex vivo intracoronary delivery of AAV2 resulted in efficient transgene expression in heart transplants, whereas intracoronary AAV9 escapes into adjacent tissues. In terms of cardiac transduction, these results suggest AAV2 as a potential vector for gene therapy in preclinical heart transplants studies, and highlight the importance of delivery route in gene transfer studies.
authors.affiliation: Transplantation Laboratory, Haartman Institute, University of Helsinki and Department of Cardiac Surgery, Heart and Lung Center, Helsinki University Central Hospital, Helsinki, Finland.
authors.forename: Alireza
authors.initials: A
authors.lastname: Raissadati
authors.forename: Janne J
authors.initials: JJ
authors.lastname: Jokinen
authors.forename: Simo O
authors.initials: SO
authors.lastname: Syrjälä
authors.forename: Mikko A I
authors.initials: MA
authors.lastname: Keränen
authors.forename: Rainer
authors.initials: R
authors.lastname: Krebs
authors.forename: Raimo
authors.initials: R
authors.lastname: Tuuminen
authors.forename: Ralica
authors.initials: R
authors.lastname: Arnaudova
authors.forename: Eeva
authors.initials: E
authors.lastname: Rouvinen
authors.forename: Andrey
authors.initials: A
authors.lastname: Anisimov
authors.forename: Jarkko
authors.initials: J
authors.lastname: Soronen
authors.forename: Katri
authors.initials: K
authors.lastname: Pajusola
authors.forename: Kari
authors.initials: K
authors.lastname: Alitalo
authors.forename: Antti I
authors.initials: AI
authors.lastname: Nykänen
authors.forename: Karl
authors.initials: K
authors.lastname: Lemström
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006321:Heart / Q000821:virology*; D016027:Heart Transplantation; D064596:Isografts; D008156:Luciferases / Q000235:genetics; D008297:Male; D009206:Myocardium / Q000378:metabolism; D051381:Rats; D019076:Transgenes / Q000235:genetics
chemical_list: D008156:Luciferases
keywords: adeno-associated virus; cardiac transduction; heart transplantation; intracoronary delivery; rat